Determination of four aminoglycoside antibiotics and spectinomycin in feed at cross-contamination level: Development and in-house validation of a LC-MS/MS method for enforcing EU Regulation.

Combating antimicrobial resistance is a top priority worldwide involving a concerted action by several high-level institutions and organisations in the health sector. To ensure that a meaningful progress is achieved, several campaigns and political initiatives have been launched targeting the health professionals, the industry, the farmers, and the general public. The Regulation (EU) 2019/4 on medicated feed contains provisions for the limitation and control of the contamination of non-target compound feed with 24 antimicrobials. The purpose of this work was to develop a reliable and effective method for the determination of four aminoglycoside antibiotics (apramycin, paromomycin, tobramycin and neomycin) and spectinomycin in feed at cross-contamination level, where an absolute lack of suitable methods was identified. Four candidate methods described in the literature failed to provide adequate recoveries of all analytes. Therefore, an in-depth investigation was carried out to identify the bottleneck variable. The optimised method was then in-house validated and showed performance features appropriate for the intended purpose. The selected compounds could be analysed by LC-MS/MS in five animal feeds with LOQs between 2.6 and 9.2 µg kg-1 for the AGs and between 28 and 86 µg kg-1 for spectinomycin. Using isotopically labelled internal standards, the recovery rates varied from 63 % to 103 % and the intermediate precision (RSDip) varied from 1.1 % to 14 %. This work represents a step forward in the reliable determination of antibiotics in compound feed as the developed method has shown to be precise and sensitive. It is expected that this method gains wide acceptance and can supplement the legislation with effective control tools for antibiotic residues.

Sustainable pig diets: partial grain replacement with former food products and its impact on meat quality.

This study investigated the effects of salty and sugary former foodstuff products (FFPs) on the quality traits and meat composition of 36 male castrated pigs (Swiss Large White breed) as well as sensory characteristics of the loins. The animals were fed three different diets for both the growing (G) and finishing (F) phases: (1) a standard diet (ST), 0% FFPs; (2) a diet with 30% of sugary FFPs (e.g., chocolate, biscuits, cakes) as a replacement for traditional ingredients (SU); and (3) a diet with 30% of salty FFPs (e.g., bread, pasta, and breadsticks) as a replacement for traditional ingredients (SA). For a comprehensive assessment of meat quality, protein and fat content in the LD were analyzed. AA and FA profile were determined both in the LD and backfat. Meat quality traits such as pH and temperature, thawing, cooking and drip losses, and shear force have been evaluated. Then, pork loins have been assessed for sensory attributes by a trained sensory panel. The SA diet decreased 20:5 n-3 levels (P < 0.001) in the muscle and 22:5 n-3 levels (P < 0.05) in both muscle and backfat but increased (P < 0.05) the ratio of mono-unsaturated to saturated fatty acids compared to the ST group. Both the SU and SA diets elevated (P < 0.001) the n-6:n-3 fatty acids ratio compared to the ST diet. Dietary treatments did not affect other meat quality traits. Regarding sensory attributes, the loin from pigs fed with SU and SA diets were sweeter (P < 0.001). Loins of SA pigs were more tender (P < 0.001), had a more intense pork aroma (P < 0.001) and had more flavor (P < 0.01) compared to ST loins. Overall, the use of FFPs affected the fatty acid profile of pork while improving the sensory quality of the loins, with no negative effects observed on the technological and nutritional quality of the meat.

Pigs are ideal species to convert food losses, also named former food products (FFPs), into animal proteins. The present study investigated the impact of incorporating sugary and salty FFPs into the diets of growing and finishing pigs on meat quality and its sensory characteristics. Our study showed that, while technological meat quality remained largely unaffected, the dietary treatments led to slight alterations of meat and backfat fatty acids profile. Moreover, a panel test for sensory analyses revealed that loins from both the sugary and salty ingredients-fed pigs were perceived as sweeter, and loins from pigs fed salty ingredients were noted for increased tenderness, intense pork aroma, and flavor. Overall, FFPs inclusion into pig diets had no detrimental effects on technological or nutritional aspects of the meat, confirming their potential use as alternative animal feed.

Preserving global land and water resources through the replacement of livestock feed crops with agricultural by-products.

While animal-source foods contribute to 16% of the global food supply and are an important protein source in human diets, their production uses a disproportionately large fraction of agricultural land and water resources. Therefore, a global comprehensive understanding of the extent to which livestock production competes directly or indirectly with food crops is needed. Here we use an agro-hydrological model combined with crop-specific yield data to investigate to what extent the replacement of some substitutable feed crops with available agricultural by-products would spare agricultural land and water resources that could be reallocated to other uses, including food crop production. We show that replacing 11-16% of energy-rich feed crops (that is, cereals and cassava) with agricultural by-products would allow for the saving of approximately 15.4-27.8 Mha of land, and 3-19.6 km3 and 74.2-137.8 km3 of blue and green water, respectively, for the growth of other food crops, thus providing a suitable strategy to reduce unsustainable use of natural resources both locally or through virtual land and water trade.

Advances in selenium supplementation: From selenium-enriched yeast to potential selenium-enriched insects, and selenium nanoparticles.

Selenium (Se) is an essential micronutrient that plays an important role in animal and human development and physiological homoeostasis. This review surveys the role of Se in the environment, plants and animal bodies, and discusses data on Se biofortification with different sources of supplementation, from inorganic to organic forms, with special focus on Se-enriched yeast (Se-yeast). Although Se-yeast remains one of the main sources of organic Se, other emerging and innovative sources are reviewed, such as Se-enriched insects and Se-nanoparticles and their potential use in animal nutrition. Se-enriched insects are discussed as an option for supplying Se in organic form to livestock diets. Se-nanoparticles are also discussed, as they represent a more biocompatible and less toxic source of inorganic Se for animal organisms, compared to selenite and selenate. We also provide up to date information on the legal framework in the EU, USA, and Canada of Se that is contained in feed additives. From the scientific evidence available in the literature, it can be concluded that among the inorganic forms, sodium selenite is still one of the main options, whereas Se-yeast remains the primary organic form. However, other potential sources such as Se-enriched insects and Se-nanoparticles are being investigated as they could potentially combine a high bioavailability and reduced Se emissions in the environment.

Compliance between Food and Feed Safety: Eight-Year Survey (2013-2021) of Aflatoxin M1 in Raw Milk and Aflatoxin B1 in Feed in Northern Italy.

Aflatoxins (AFs) are fungal metabolites that are found in feed and food. When ruminants eat feed contaminated with aflatoxin B1 (AFB1), it is metabolised and aflatoxin M1 (AFM1) is excreted in the milk. Aflatoxins can result in hepatotoxic, carcinogenic, and immunosuppressive effects. The European Union thus set a low threshold limit (50 ng/L) for presence of AFM1 in milk. This was in view of its possible presence also in dairy products and that quantification of these toxins is mandatory for milk suppliers. In the present study, a total of 95,882 samples of whole raw milk, collected in northern Italy between 2013 and 2021, were evaluated for presence of AFM1 using an ELISA (enzyme-linked immunosorbent assay) method. The study also evaluated the relationship between feed materials collected from the same farms in the same area during the same period (2013-2021) and milk contamination. Only 667 milk samples out of 95,882 samples analysed (0.7%) showed AFM1 values higher than the EU threshold limit of 50 ng/L. A total of 390 samples (0.4%) showed values between 40 and 50 ng/L, thus requiring corrective action despite not surpassing the regulatory threshold. Combining feed contamination and milk contamination data, some feedingstuffs seem to be more effective in defying potential carryover of AFs from feed to milk. Combining the results, it can be concluded that a robust monitoring system that covers both feed, with a special focus on high risk/sentinel matrices, and milk is essential to guarantee high quality and safety standards of dairy products.

Packaging contaminants in former food products: Using Fourier Transform Infrared Spectroscopy to identify the remnants and the associated risks.

Food waste and feed-food competition can be reduced by replacing traditional feed ingredients such as cereals, with former food products (FFPs) in livestock diets. These foodstuffs, initially intended for human consumption, are recovered, mechanically unpacked, and then ground. Despite this simple and inexpensive treatment, packaging contaminants (remnants) are often unavoidable in the final product. To maximize the exploitation of FFPs and to minimize the associated risks, packaging remnants need to be quantified and characterized. This study tested the efficacy of the Fourier Transform Infrared Spectroscopy coupled with an optical microscope (µFT-IR) in identifying packaging remnants in 17 FFP samples collected in different geographical areas. After a visual sorting procedure, presumed packaging remnants were analyzed by µFT-IR. The results showed significant differences (p < 0.05) between the FFPs in terms of the total number of foreign particles found (plastics, cellulose and aluminum remnants, ranging from 4 to 19 particles per 20 g fresh matter), and also regarding the number of cellulose and aluminum particles. These data clearly demonstrate the need for sensitive instruments that can characterize the potential contaminants in the FFPs. This would then help to reduce the overestimation of undesirable contaminants typical of simple visual sorting, which is currently the most common method.

E-Nose Technology for Mycotoxin Detection in Feed: Ready for a Real Context in Field Application or Still an Emerging Technology?

Mycotoxin risk in the feed supply chain poses a concern to animal and human health, economy, and international trade of agri-food commodities. Mycotoxin contamination in feed and food is unavoidable and unpredictable. Therefore, monitoring and control are the critical points. Effective and rapid methods for mycotoxin detection, at the levels set by the regulations, are needed for an efficient mycotoxin management. This review provides an overview of the use of the electronic nose (e-nose) as an effective tool for rapid mycotoxin detection and management of the mycotoxin risk at feed business level. E-nose has a high discrimination accuracy between non-contaminated and single-mycotoxin-contaminated grain. However, the predictive accuracy of e-nose is still limited and unsuitable for in-field application, where mycotoxin co-contamination occurs. Further research needs to be focused on the sensor materials, data analysis, pattern recognition systems, and a better understanding of the needs of the feed industry for a safety and quality management of the feed supply chain. A universal e-nose for mycotoxin detection is not realistic; a unique e-nose must be designed for each specific application. Robust and suitable e-nose method and advancements in signal processing algorithms must be validated for specific needs.

Cold-Pressed Oilseed Cakes as Alternative and Sustainable Feed Ingredients: A Review.

Due to the increasing demand for alternative protein feed ingredients, the utilization of oilseed by-products in animal nutrition has been sought as a promising solution to ensure cheap and environmentally sustainable feedstuffs. This review aimed to summarize the nutritional value of six cold-pressed cakes (rapeseed, hempseed, linseed, sunflower seed, camelina seed, and pumpkin seed) and the effects of their inclusion in diet for ruminant, pig, and poultry on nutrient digestibility, growth and productive performance, and quality of the products. The presented results indicated that these unconventional feed ingredients are a good protein and lipid source and have a balanced amino acid and fatty acid profile. However, contradictory results of animal production performances can be found in the literature depending on the cake type and chemical composition, dietary inclusion level, animal category, and trial duration. Due to the substantial amount of essential fatty acid, these cakes can be efficiently used in the production of animal products rich in n-3 and n-6 polyunsaturated fatty acids. However, the utilization of cakes in pig and poultry nutrition is limited because of the presence of antinutritive factors that can deteriorate feed intake and nutrient utilization.

An Eight-Year Survey on Aflatoxin B1 Indicates High Feed Safety in Animal Feed and Forages in Northern Italy.

Aflatoxins (AFs) remain the main concern for the agricultural and dairy industries due to their effects on the performances and quality of livestock production. Aflatoxins are always unavoidable and should be monitored. The objective of this paper is to bring to light a significant volume of data on AF contamination in several animal feed ingredients in Northern Italy. The Regional Breeders Association of Lombardy has been conducting a survey program to monitor mycotoxin contamination in animal feeds, and in this paper, we present data relating to AFB1 contamination. In most cases (95%), the concentrations were low enough to ensure compliance with the European Union's (EU's) maximum admitted levels for animal feed ingredients. However, the data show a high variability in AF contamination between different matrices and, within the same matrix, a high variability year over year. High levels of AFs were detected in maize and cotton, especially in the central part of the second decade of this century, i.e., 2015-2018, which has shown a higher risk of AF contamination in feed materials in Northern Italy. Variability due to climate change and the international commodity market affect future prospects to predict the presence of AFs. Supplier monitoring and control and reduced buying of contaminated raw materials, as well as performing analyses of each batch, help reduce AF spread.

Fate of Salmonella enterica Typhimurium and Listeria monocytogenes in Black Soldier Fly (Hermetia illucens) Larvae Reared on Two Artificial Diets.

Ensuring food security is one of the main challenges facing the world over the next 30 years. There is, thus, an urgent need to significantly increase the supply of sustainable protein that can be transformed into animal feed. Proteins from insects offer a valuable alternative. This article presents the results of challenge tests conducted to investigate the dynamics of the microbial load of Salmonella enterica Typhimurium and Listeria monocytogenes in black soldier fly (Hermetia illucens) larvae grown on contaminated substrates. Four separate challenge tests were performed on two substrates: the Gainesville diet and a homemade diet. The challenge test procedure was carried out in accordance with ISO/DIS 20976-2 (under development). The results of this study show that, when grown on contaminated substrates, BSF larvae do not eliminate Salmonella Typhimurium or L. monocytogenes, but can reduce their microbial load. Sanitation processes downstream of the breeding of BSF larvae are, however, required to reduce the microbiological risks of this novel food.

Characterization of a Bacillus cereus strain associated with a large feed-related outbreak of severe infection in pigs.

AIMS: Bacillus cereus is often responsible for foodborne diseases and both local and systemic infections in humans. Cases of infection in other mammals are rather rare. In this study, we report a B. cereus feed-related outbreak that caused the death of 6234 pigs in Italy. METHODS AND RESULTS: Massive doses of a Gram-positive, spore-forming bacterium were recovered from the animal feed, faeces of survived pigs and intestinal content of dead ones. The B. cereus MM1 strain was identified by MALDI-TOF MS and typified by RAPD-PCR. The isolate was tested for the production of PC-PLC, proteases, hemolysins and biofilm, for motility, as well as for the presence of genes encoding tissue-degrading enzymes and toxins. Antimicrobial resistance and pathogenicity in Galleria mellonella larvae were also investigated. Our results show that the isolated B. cereus strain is swimming-proficient, produces PC-PLC, proteases, hemolysins, biofilm and carries many virulence genes. The strain shows high pathogenicity in G. mellonella larvae. CONCLUSIONS: The isolated B. cereus strain demonstrates an aggressive profile of pathogenicity and virulence, being able to produce a wide range of determinants potentially hazardous to pigs' health. SIGNIFICANCE AND IMPACT OF STUDY: This study highlights the proficiency of B. cereus to behave as a devastating pathogen in swine if ingested at high doses and underlines that more stringent quality controls are needed for livestock feeds and supplements.

Sweet vs. Salty Former Food Products in Post-Weaning Piglets: Effects on Growth, Apparent Total Tract Digestibility and Blood Metabolites.

Former food products (FFPs) have a great potential to replace conventional feed ingredients. This study aimed to investigate the possibility to partially replace standard ingredients with two different types of FFPs: bakery (FFPs-B) or confectionary (FFPs-C) FFPs and their effects on growth performances, feed digestibility and metabolic status in post-weaning piglets. Thirty-six post-weaning piglets were randomly assigned to three experimental diets (n = 12 per diet) for 42 days: a standard diet (CTR), a diet where 30% of standard ingredients were replaced by confectionary FFPs (FFPs-C) and a diet where 30% of standard ingredients were replaced by bakery FFPs (FFPs-B). Individual body weight and fecal dry matter were measured weekly. Feed intake (FI) was determined daily. Average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR) were calculated. Fecal samples were collected daily for three days/week to determine apparent total tract digestibility of dry matter (ATTD). At day 0, 21 and 42, blood samples were collected from all the piglets. No significant differences (p > 0.05) between groups were found in growth performances and metabolic profile. However, ATTD in FFPs-B group was lower (p < 0.05) compared to the CTR group at the end of the experiment. This study confirmed the possibility to formulate homogeneous diets integrated with 30% of both categories of FFPs. Further investigations are needed to clarify the effects of bakery former food products on the digestibility of the diet.

Integrated Mycotoxin Management System in the Feed Supply Chain: Innovative Approaches.

Exposure to mycotoxins is a worldwide concern as their occurrence is unavoidable and varies among geographical regions. Mycotoxins can affect the performance and quality of livestock production and act as carriers putting human health at risk. Feed can be contaminated by various fungal species, and mycotoxins co-occurrence, and modified and emerging mycotoxins are at the centre of modern mycotoxin research. Preventing mould and mycotoxin contamination is almost impossible; it is necessary for producers to implement a comprehensive mycotoxin management program to moderate these risks along the animal feed supply chain in an HACCP perspective. The objective of this paper is to suggest an innovative integrated system for handling mycotoxins in the feed chain, with an emphasis on novel strategies for mycotoxin control. Specific and selected technologies, such as nanotechnologies, and management protocols are reported as promising and sustainable options for implementing mycotoxins control, prevention, and management. Further research should be concentrated on methods to determine multi-contaminated samples, and emerging and modified mycotoxins.

Soybean Molasses in Animal Nutrition.

Concerning the increasing global demand for food and accumulation of huge amounts of biomass waste from the agro-food industry whose manipulation is usually inadequate, the potential of livestock to convert by-products as alternative feed ingredients into valuable proteins has been proposed as an outstanding option. Soybean molasses present a by-product of soybean protein concentrate production with low commercial cost but high nutritive and functional value. It is a rich source of soluble carbohydrates in the form of sugars and soybean phytochemicals. Therefore, this paper provides a review of published works about the production of soybean molasses, chemical composition, and nutritive value. In addition, the possibility of the application of soybean molasses in animal nutrition as a pelleting aid and functional feed ingredient is also discussed. Special attention is devoted to the influence of the inclusion of soybean molasses in the diets for ruminants, non-ruminants, and aquaculture on animal performance and health.

The Contribution of Dietary Magnesium in Farm Animals and Human Nutrition.

Magnesium (Mg) is a mineral that plays an essential role as cofactor of more than 300 enzymes. Mg in farm animals' and human nutrition is recommended to avoid Mg deficiency, ensure adequate growth and health maintenance. Mg supplementation above the estimated minimum requirements is the best practice to improve farm animals' performances (fertility and yield) and food products' quality, since the performance of farm animals has grown in recent decades. Mg supplementation in pigs increases meat quality and sows' fertility; in poultry, it helps to avoid deficiency-related health conditions and to improve meat quality and egg production by laying hens; in dairy cows, it serves to avoid grass tetany and milk fever, two conditions related to hypomagnesaemia, and to support their growth. Thus, Mg supplementation increases food products' quality and prevents Mg deficiency in farm animals, ensuring an adequate Mg content in animal-source food. These latter are excellent Mg sources in human diets. Sub-optimal Mg intake by humans has several implications in bone development, muscle function, and health maintenance. This review summarizes the main knowledge about Mg in farm animals and in human nutrition.

Magnesium in Obesity, Metabolic Syndrome, and Type 2 Diabetes.

Magnesium (Mg2+) deficiency is probably the most underestimated electrolyte imbalance in Western countries. It is frequent in obese patients, subjects with type-2 diabetes and metabolic syndrome, both in adulthood and in childhood. This narrative review aims to offer insights into the pathophysiological mechanisms linking Mg2+ deficiency with obesity and the risk of developing metabolic syndrome and type 2 diabetes. Literature highlights critical issues about the treatment of Mg2+ deficiency, such as the lack of a clear definition of Mg2+ nutritional status, the use of different Mg2+ salts and dosage and the different duration of the Mg2+ supplementation. Despite the lack of agreement, an appropriate dietary pattern, including the right intake of Mg2+, improves metabolic syndrome by reducing blood pressure, hyperglycemia, and hypertriglyceridemia. This occurs through the modulation of gene expression and proteomic profile as well as through a positive influence on the composition of the intestinal microbiota and the metabolism of vitamins B1 and D.

Gravimetric quantitative validation of botanic impurities in feed.

BACKGROUND: Harmful botanical impurities may contaminate feed and feed materials and be a potential danger to animal or human health, or to the environment. The aim of this study was to establish rapid and sensitive methods that can be used in routine official controls to determine botanical impurities such as Datura stramonium, Ricinus communis, Crotaliaria spp., and Ambrosia spp. in animal feed and raw materials. Claviceps sclerotia were also detected in cereals, due to the similarities of the targets and the analytical procedure. Regulation (EU) 625/2017, which replaces Reg. 2004/882/EC, states that EU member states should conduct official controls in assessed and accredited laboratories and that the analytical methods must be validated before use by considering parameters such as specificity, precision, recovery, and measurement uncertainly. RESULTS AND CONCLUSION: The results demonstrate that all of the methods tested are suitable for the official quantitative analyses required by EU official legislation. © 2020 Society of Chemical Industry.

Going to the roots of reduced magnesium dietary intake: A tradeoff between climate changes and sources.

Magnesium is essential in plants where it is associated with chlorophyll pigments and serves as a cofactor of enzymes implicated in photosynthesis and metabolism. It is an essential nutrient for animals, involved in hundreds metabolic reaction and crucial for the biological activity of ATP. Not surprisingly, magnesium deficiency is detrimental for the health of plants and animals. In humans, subclinical magnesium deficiency is common and generates chronic inflammation, which is the common denominator of a wide range of mental and physical health problems from metabolic diseases to cognitive impairment, from osteopenia and sarcopenia to depression. It is ascertained that magnesium content in fruits and vegetables dropped in the last fifty years, and about 80% of this metal is lost during food processing. As a consequence, a large percentage of people all over the world does not meet the minimum daily magnesium requirement. In this scoping review, we summarize how agronomic and environmental factors, including global warming, affect magnesium content and availability in the soil and, consequently, in the food chain, with the aim of attracting the interest of botanists, agronomists, animal and human nutritionists and physicians to work on a strategy that grants adequate magnesium intake for everybody.

Comparative Proteomics of Milk Fat Globule Membrane (MFGM) Proteome across Species and Lactation Stages and the Potentials of MFGM Fractions in Infant Formula Preparation.

Milk is a lipid-in-water emulsion with a primary role in the nutrition of newborns. Milk fat globules (MFGs) are a mixture of proteins and lipids with nutraceutical properties related to the milk fat globule membrane (MFGM), which protects them, thus preventing their coalescence. Human and bovine MFGM proteomes have been extensively characterized in terms of their formation, maturation, and composition. Here, we review the most recent comparative proteomic analyses of MFGM proteome, above all from humans and bovines, but also from other species. The major MFGM proteins are found in all the MFGM proteomes of the different species, although there are variations in protein expression levels and molecular functions across species and lactation stages. Given the similarities between the human and bovine MFGM and the bioactive properties of MFGM components, several attempts have been made to supplement infant formulas (IFs), mainly with polar lipid fractions of bovine MFGM and to a lesser extent with protein fractions. The aim is thus to narrow the gap between human breast milk and cow-based IFs. Despite the few attempts made to date, supplementation with MFGM proteins seems promising as MFGM lipid supplementation. A deeper understanding of MFGM proteomes should lead to better results.

Multivariate image analysis for the rapid detection of residues from packaging remnants in former foodstuff products (FFPs) - a feasibility study.

From a circular economy perspective, feeding livestock with food leftovers or former foodstuff products (FFPs) could be an effective option aimed at exploiting food leftover resources and reducing food losses. FFPs are valuable energy sources, characterised by a beneficial starch/sugar content, and also fats. However, besides these nutritional aspects, safety is a key concern given that FFPs are generally derived from packaged food. Packaging materials, such as plastics and paper, are not accepted as a feed ingredient which means that residues should be rigorously avoided. A sensitive and objective detection method is thus essential for an accurate risk evaluation throughout the former food production chain. To this end, former food samples were collected in processing plants of two different European countries and subjected to multivariate analysis of red, green, and blue (RGB) microscopic images, in order to evaluate the possible application of this non-destructive technique for the rapid detection of residual particles from packaging materials. Multivariate Image Analysis (MIA) was performed on single images at the pixel level, which essentially consisted in an exploratory analysis of the image data by means of Principal Component Analysis, which highlighted the differences between packaging and foodstuff particles, based on their colour. The whole dataset of images was then analysed by means of a multivariate data dimensionality reduction method known as the colourgrams approach, which identified clusters of images sharing similar features and also highlighted outlier images due to the presence of packaging particles. The results obtained in this feasibility study demonstrated that MIA is a promising tool for a rapid automated method for detecting particles of packaging materials in FFPs.